Revision 4

#67840Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IF-F, IF-IC, eCLIP

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

70

Source/Isotype:

Rabbit IgG

UniProt ID:

#P35637

Entrez-Gene Id:

2521

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunofluorescence (Frozen) 1:800 - 1:3200
Immunofluorescence (Immunocytochemistry) 1:800 - 1:3200
eCLIP 1:200
For more information about the RBP-eCLIP service please visit Eclipsebio.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier-free (BSA and azide free) version of this product see product #48647.

Specificity / Sensitivity

FUS/TLS (E3O8I) Rabbit mAb recognizes endogenous levels of total FUS/TLS protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gln146 of human FUS/TLS protein.

Background

FUS/TLS (fused in sarcoma/translocated in liposarcoma) was initially identified by investigators as a component of fusion proteins found in a variety of cancers, such as myxoid liposarcoma, acute myeloid leukemia, and Ewing’s tumor (1). FUS/TLS fusion with the DNA-binding domain of transcription activators, such as CHOP and ERG, leads to aberrant transcription of target genes that is thought by researchers to lead to tumor development (1-5). FUS/TLS is involved in a wide range of RNA processing events, such as pre-mRNA splicing, mRNA transcription, and miRNA processing (1,6). In addition to its role in RNA metabolism, FUS/TLS maintains genomic stability and co-regulates gene expression by interacting with various transcription factors such as nuclear receptors, YB-1, p65 subunit of NF-κB, TFIID, and RUNX2 (1,6,7). More recently, researchers have found several mutations of FUS/TLS in ALS (amyotrophic lateral sclerosis) and FTLD (frontotemporal lobar degeneration) patients that causes cytoplasmic mislocalization of FUS/TLS (6,8-12).

  1. Yang, S. et al. (2010) Int J Biochem Cell Biol 42, 1408-11.
  2. Crozat, A. et al. (1993) Nature 363, 640-4.
  3. Rabbitts, T.H. et al. (1993) Nat Genet 4, 175-80.
  4. Law, W.J. et al. (2006) Brief Funct Genomic Proteomic 5, 8-14.
  5. Prasad, D.D. et al. (1994) Oncogene 9, 3717-29.
  6. Lagier-Tourenne, C. et al. (2010) Hum Mol Genet 19, R46-64.
  7. Baechtold, H. et al. (1999) J Biol Chem 274, 34337-42.
  8. Hewitt, C. et al. (2010) Arch Neurol 67, 455-61.
  9. Vance, C. et al. (2009) Science 323, 1208-11.
  10. Van Langenhove, T. et al. (2010) Neurology 74, 366-71.
  11. Da Cruz, S. and Cleveland, D.W. (2011) Curr Opin Neurobiol 21, 904-19.
  12. Hock, E.M. et al. (2018) Cell Rep 24, 987-1000.e7.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry) eCLIP: eCLIP

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 4
#67840

FUS/TLS (E3O8I) Rabbit mAb

Western Blotting Image 1: FUS/TLS (E3O8I) Rabbit mAb Expand Image
Western blot analysis of extracts from mouse brain tissue and SH-SY5Y cells using FUS/TLS (E3O8I) Rabbit mAb (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
Immunoprecipitation Image 1: FUS/TLS (E3O8I) Rabbit mAb Expand Image
Immunoprecipitation of FUS/TLS protein from mouse brain tissue extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is FUS/TLS (E3O8I) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Immunofluorescence Image 1: FUS/TLS (E3O8I) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of mouse colon using FUS/TLS (E3O8I) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Sections were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 2: FUS/TLS (E3O8I) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of mouse liver using FUS/TLS (E3O8I) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Sections were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 3: FUS/TLS (E3O8I) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of mouse retina using FUS/TLS (E3O8I) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Sections were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 1: FUS/TLS (E3O8I) Rabbit mAb Expand Image
Immunofluorescent analysis of SH-SY5Y cells, untreated (left) or exposed to hypertonic stress (400 mM sorbitol, 2 hr; right), using FUS/TLS (E3O8I) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue). Cytoplasmic translocation is consistent with the findings reported by Hock, E.M. et al. (12), confirming specificity of the antibody for FUS/TLS.
eCLIP Image 1: FUS/TLS (E3O8I) Rabbit mAb Expand Image
Enhanced cross-linking and immunoprecipitation (eCLIP) was performed with RNA from K-562 cells and FUS/TLS (E3O8I) Rabbit mAb using a protocol based on the RBP-eCLIP method from Eclipsebio. The figure shows binding across the FUS transcript. Data is kindly provided by the laboratory of Dr. Gene Yeo and used with permission.